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Three-Dimensional Reconstruction of Neurovascular Network in Whole Mount Preparations and Thick-Cut Transverse Sections of Mouse Urinary Bladder

PURPOSE: Proper functional and structural integrity of nervous and vascular system in urinary bladder plays an important role in normal bladder function and the disruption of these structures is known to be related to lower urinary tract symptoms. Here, we present an immunohistochemical staining met...

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Autores principales: Nguyen, Nhat Minh, Song, Kang-Moon, Choi, Min-Ji, Ghatak, Kalyan, Limanjaya, Anita, Kwon, Mi-Hye, Chung, Doo Yong, Ock, Jiyeon, Yin, Guo Nan, Park, Chang-Shin, Suh, Jun-Kyu, Ryu, Ji-Kan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Korean Society for Sexual Medicine and Andrology 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7752506/
https://www.ncbi.nlm.nih.gov/pubmed/31496149
http://dx.doi.org/10.5534/wjmh.190089
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author Nguyen, Nhat Minh
Song, Kang-Moon
Choi, Min-Ji
Ghatak, Kalyan
Limanjaya, Anita
Kwon, Mi-Hye
Chung, Doo Yong
Ock, Jiyeon
Yin, Guo Nan
Park, Chang-Shin
Suh, Jun-Kyu
Ryu, Ji-Kan
author_facet Nguyen, Nhat Minh
Song, Kang-Moon
Choi, Min-Ji
Ghatak, Kalyan
Limanjaya, Anita
Kwon, Mi-Hye
Chung, Doo Yong
Ock, Jiyeon
Yin, Guo Nan
Park, Chang-Shin
Suh, Jun-Kyu
Ryu, Ji-Kan
author_sort Nguyen, Nhat Minh
collection PubMed
description PURPOSE: Proper functional and structural integrity of nervous and vascular system in urinary bladder plays an important role in normal bladder function and the disruption of these structures is known to be related to lower urinary tract symptoms. Here, we present an immunohistochemical staining method that delineates neurovascular structures in the mouse urinary bladder by using immunohistochemical staining with three-dimensional reconstruction. MATERIALS AND METHODS: The urinary bladder was harvested from 8-week-old C57BL/6 male mouse. Lamina propria and detrusor muscle layer were dissected for whole mount staining, and thick-cut (60-µm) sections were prepared for full-thickness bladder staining. Immunofluorescent staining of bladder tissue was performed with antibodies against CD31 (an endothelial cell marker), smooth muscle α-actin (a smooth muscle cell marker), NG2 (a pericyte marker), and βIII-tubulin (a neuronal marker). We reconstructed three-dimensional images of bladder neurovascular system from stacks of two-dimensional images. RESULTS: Three-dimensional images obtained from thick-cut sections clearly provided good anatomic information about neurovascular structures in the three layers of bladder, such as urothelium, lamina propria, and detrusor muscle layer. Whole mount images of lamina propria and detrusor muscle layer also clearly delineated spatial relationship between nervous and vascular systems. The microvessel density was higher in the lamina propria than in the detrusor muscle layer. Nerve fibers were evenly innervated into the lamina propria and detrusor muscle. CONCLUSIONS: This study provides comprehensive insight into three-dimensional neurovascular structures of mouse urinary bladder. Our technique may constitute a standard tool to evaluate pathologic changes in a variety of urinary bladder diseases.
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spelling pubmed-77525062021-01-05 Three-Dimensional Reconstruction of Neurovascular Network in Whole Mount Preparations and Thick-Cut Transverse Sections of Mouse Urinary Bladder Nguyen, Nhat Minh Song, Kang-Moon Choi, Min-Ji Ghatak, Kalyan Limanjaya, Anita Kwon, Mi-Hye Chung, Doo Yong Ock, Jiyeon Yin, Guo Nan Park, Chang-Shin Suh, Jun-Kyu Ryu, Ji-Kan World J Mens Health Original Article PURPOSE: Proper functional and structural integrity of nervous and vascular system in urinary bladder plays an important role in normal bladder function and the disruption of these structures is known to be related to lower urinary tract symptoms. Here, we present an immunohistochemical staining method that delineates neurovascular structures in the mouse urinary bladder by using immunohistochemical staining with three-dimensional reconstruction. MATERIALS AND METHODS: The urinary bladder was harvested from 8-week-old C57BL/6 male mouse. Lamina propria and detrusor muscle layer were dissected for whole mount staining, and thick-cut (60-µm) sections were prepared for full-thickness bladder staining. Immunofluorescent staining of bladder tissue was performed with antibodies against CD31 (an endothelial cell marker), smooth muscle α-actin (a smooth muscle cell marker), NG2 (a pericyte marker), and βIII-tubulin (a neuronal marker). We reconstructed three-dimensional images of bladder neurovascular system from stacks of two-dimensional images. RESULTS: Three-dimensional images obtained from thick-cut sections clearly provided good anatomic information about neurovascular structures in the three layers of bladder, such as urothelium, lamina propria, and detrusor muscle layer. Whole mount images of lamina propria and detrusor muscle layer also clearly delineated spatial relationship between nervous and vascular systems. The microvessel density was higher in the lamina propria than in the detrusor muscle layer. Nerve fibers were evenly innervated into the lamina propria and detrusor muscle. CONCLUSIONS: This study provides comprehensive insight into three-dimensional neurovascular structures of mouse urinary bladder. Our technique may constitute a standard tool to evaluate pathologic changes in a variety of urinary bladder diseases. Korean Society for Sexual Medicine and Andrology 2021-01 2019-08-27 /pmc/articles/PMC7752506/ /pubmed/31496149 http://dx.doi.org/10.5534/wjmh.190089 Text en Copyright © 2021 Korean Society for Sexual Medicine and Andrology http://creativecommons.org/licenses/by-nc/4.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Article
Nguyen, Nhat Minh
Song, Kang-Moon
Choi, Min-Ji
Ghatak, Kalyan
Limanjaya, Anita
Kwon, Mi-Hye
Chung, Doo Yong
Ock, Jiyeon
Yin, Guo Nan
Park, Chang-Shin
Suh, Jun-Kyu
Ryu, Ji-Kan
Three-Dimensional Reconstruction of Neurovascular Network in Whole Mount Preparations and Thick-Cut Transverse Sections of Mouse Urinary Bladder
title Three-Dimensional Reconstruction of Neurovascular Network in Whole Mount Preparations and Thick-Cut Transverse Sections of Mouse Urinary Bladder
title_full Three-Dimensional Reconstruction of Neurovascular Network in Whole Mount Preparations and Thick-Cut Transverse Sections of Mouse Urinary Bladder
title_fullStr Three-Dimensional Reconstruction of Neurovascular Network in Whole Mount Preparations and Thick-Cut Transverse Sections of Mouse Urinary Bladder
title_full_unstemmed Three-Dimensional Reconstruction of Neurovascular Network in Whole Mount Preparations and Thick-Cut Transverse Sections of Mouse Urinary Bladder
title_short Three-Dimensional Reconstruction of Neurovascular Network in Whole Mount Preparations and Thick-Cut Transverse Sections of Mouse Urinary Bladder
title_sort three-dimensional reconstruction of neurovascular network in whole mount preparations and thick-cut transverse sections of mouse urinary bladder
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7752506/
https://www.ncbi.nlm.nih.gov/pubmed/31496149
http://dx.doi.org/10.5534/wjmh.190089
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